双酚 AF（BPAF）是一种新兴的内分泌干扰化学物质（EDC），作为双酚 A 的主要替代品之一，在环境中普遍存在。EDC的性别特异性效应已被广泛报道，并与在不同暴露窗口期间性别二态激素代谢模式和相关基因表达紧密相关，但我们对这些机制的理解仍然有限。本研究以10 μg/L 的环境相关浓度暴露成年斑马鱼28天，通过RNA测序（RNA-seq）和Ingenuity Pathway Analysis（IPA）分别研究了雄性和雌性鱼的肝脏在长期BPAF 暴露下的全局转录网络，连接了BPAF 长期暴露对分子反应的性别依赖性毒性。结果显示，在雄性鱼中检测到的差异表达基因（DEGs）较雌性鱼（195）多（811个），而在雄性中，精子发生是富集度最高的Gene Ontology（GO）功能分类，而在雌性中，基因表达的昼夜节律调节是富集度最高的GO 术语。选择性DEGs的表达水平经过qRT-PCR例行验证，与RNA-seq数据中的转录变化一致。IPA的因果网络分析认为，在雄性中，BPAF的不良结果包括肝损伤、细胞凋亡、器官炎症和肝癌，与RNA-seq检测到的几个关键DEG的调控相关，可能与上游调控分子ifnα、yap1和ptger2的激活有关；在雌性中，上游调节因子hif1α、ifng和igf1的抑制导致了几个关键DEG的下调表达，可能参与了BPAF对雌性的影响。此外，BPAF 暴露改变了雄性和雌性肝脏的组织结构，并抑制了抗氧化能力。总之，本研究揭示了BPAF对鱼类肝脏性别依赖效应的不同调控网络，同时这些BPAF 暴露下检测到的DEGs可能作为评估环境EDC暴露后的性别特异性肝毒性的潜在生物标志物。版权所有©2023年作者。由Elsevier Inc.发表。保留所有权利。

Bisphenol AF (BPAF) is an emerging endocrine-disrupting chemical (EDC) prevalent in the environment as one of the main substitutes for bisphenol A. Sex-specific effects of EDCs have been commonly reported and closely linked to sexually dimorphic patterns of hormone metabolism and related gene expression during different exposure windows, but our understanding of these mechanisms is still limited. Here, following 28-day exposure of adult zebrafish to an environmentally relevant concentration of BPAF at 10 μg/L, the global transcriptional networks applying RNA sequencing (RNA-seq) and Ingenuity Pathway Analysis (IPA) were respectively investigated in the male and female fish liver, connecting the sex-dependent toxicity of the long-term exposure of BPAF to molecular responses. As a result, more differentially expressed genes (DEGs) were detected in males (811) than in females (195), and spermatogenesis was the most enriched Gene Ontology (GO) functional classification in males, while circadian regulation of gene expression was the most enriched GO term in females. The expression levels of selected DEGs were routinely verified using qRT-PCR, which showed consistent alterations with the transcriptional changes in RNA-seq data. The causal network analysis by IPA suggested that the adverse outcomes of BPAF in males including liver damage, apoptosis, inflammation of organ, and liver carcinoma, associated with the regulation of several key DEGs detected in RNA-seq, could be linked to the activation of upstream regulatory molecules ifnα, yap1, and ptger2; while, the inhibition of upstream regulators hif1α, ifng, and igf1, leading to the down-regulated expression of several key DEGs, might be involved in BPAF's effects in females. Furthermore, BPAF exposure altered hepatic histological structure and inhibited antioxidant capability in both male and female livers. Overall, this study revealed different regulation networks involved in the sex-dependent effects of BPAF on the fish liver, and these detected DEGs upon BPAF exposure might be used as potential biomarkers for further assessing sex-specific hepatotoxicity following environmental EDC exposure.Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.